Genetic assessment of apple germplasm in Bosnia and Herzegovina using microsatellite and morphologic markers

Traditional apple cultivars from Bosnia and Herzegovina (B&H), potentially diverse due to specific geographic location and history of the country, represent a possible source of valuable traits for future breeding efforts and sustainable fruit growing. A total of 39 accessions, 24 traditional B&H cultivars and 15 modern international cultivars, maintained at the ex situ apple collection “Srebrenik” in Northeast Bosnia were, investigated using 10 SSR (simple sequence repeats) markers and 23 morphologic characteristics. All the used primer pairs manage to amplify clearly distinguishable and highly polymorphic SSR alleles, in average 10.4 alleles per locus. More than two different alleles per locus were detected for seven accessions (five traditional B&H cultivars and two international cultivars). Forty one unique alleles were exclusively present within the B&H cultivars, while seven unique alleles were only detected within international cultivars. The differentiation between traditional B&H and international cultivars (Fst = 0.060; P < 0.0001) was significant, also confirmed by analyses of molecular variance (AMOVA) (f_CT = 0.092; P < 0.001). Cluster analyses of 39 apple accessions, based on 10 SSR loci, revealed that only two traditional B&H cultivars grouped tightly with international cultivars (Ljepocvjetka and Bobovec Jon), while the rest formed separate clusters. Multivariate analyses of variance (MANOVA), nonparametric multivariate analyses of variance (NPMANOVA) and analyses of similarity (ANOSIM) showed statistically significant difference in morphologic characteristics between traditional B&H cultivars and the international cultivars. Cluster analyses of 39 apple accessions, based on the morphologic data, displayed less differentiation between traditional and international accessions, in comparison to the cluster analyses based on molecular data. No correlation between the molecular and morphologic data set was detected using the Mantel test. Many of the morphologic characteristics which have been analyzed in this study have significant commercial importance, we can assume that unlike the microsatellites these traits have been under agronomic selection pressure.